Next Generation Disease Screening using Nanosensors'; Highly multiplexed label-free detection of soulble proteins and miRNA in biofluids

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There is an enormous need for analytical methods that can achieve simultaneous detection of multiple soluble proteins and miRNA in complex biological fluids. A technology that can achieve this holds the promise of far-reaching impact in multiple healthcare grand challenges ranging from neurodegenerative disease to several major cancers. This project aims to develop a multiplexed label-free detection of soluble proteins and miRNA in biofluids based on the Oxford Nanopore MinION. The MinION is an established platform capable of high-throughput sequencing capable probing 512 channels simultaneously. Via customisation and use of molecular carriers, we believe that such a platform can be used for the detection of up to 44 (256) protein and miRNA biomarkers simultaneously in a label-free manner. The molecular carriers will not only enable efficient transport and detection of the biomarker to the nanopore but will also incorporate a unique DNA barcode identifier consisting of 4 bases which can be used to confirm concentration and presence of a particular biomarker. Each carrier will contain a complementary sequence for the detection of miRNA or an aptamer sequence for detection of protein. As a proof of concept, we have selected 2 key proteins linked to a major neurodegenerative disease (Parkinson's, Alzheimer's) and 10 miRNA sequences which are either up or down regulated in patients. The developed method is universal and if successful this pilot work will build the basis for a general approach for the detection of proteins and small molecules such as miRNA and neurotransmitters in complex unmodified samples.